Elucidating rotaviral condensate structure and function for future antiviral targeting

阐明轮状病毒凝聚物的结构和功能以用于未来的抗病毒靶向

• 批准号: 2889083
• 金额: --
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2889083
• 关键词: Elucidating; rotaviral; condensate; structure; function

Nearly all known human viruses replicate within specialised compartments in cells called viroplasms. The phase-separating nature of these structures makes them promising antiviral targets in rotaviruses; combining high-throughputmicrofluidics approaches with biophysics and molecular virology will offer unique insights into their behaviour. For example, NSP5 associated with condensate formation phosphorylates throughout infection. Microfluidics would allowinvestigations into the effects of phosphorylation on phase boundaries; alongside in vivo studies, this could reveal why viroplasms solidify over time. The role of viroplasm-associated RNA in phase separation and particle assembly alsoremains unclear. While the disordered state of these molecules makes structure prediction challenging, reverse genetics can be used to rescue viruses and their RNA. By detecting differential FRET emissions inside droplets and comparingwith known structures, structural changes in viral RNA under various conditions could be identified. Additionally high-throughput screening can identify potential antivirals that specifically aggregate into and disrupt viral condensates. Abetter understanding of viral phase separation, along with the skillset I will have acquired from this project, could then be applied to other infectious systems. This is of particular importance when facing of a global crisis caused by antiviral and antimicrobial resistance.

几乎所有已知的人类病毒都在称为病毒囊的细胞中的专门区室中复制。这些结构的相分离性质使它们成为轮状病毒中有前途的抗病毒靶点;将高通量微流体方法与生物物理学和分子病毒学相结合将为其行为提供独特的见解。例如，与冷凝物形成相关的NSP 5在整个感染过程中磷酸化。微流体技术将允许研究磷酸化对相边界的影响;与体内研究一起，这可能揭示为什么病毒随着时间的推移而固化。病毒质相关RNA在相分离和粒子组装中的作用也尚不清楚。虽然这些分子的无序状态使结构预测具有挑战性，但反向遗传学可以用来拯救病毒及其RNA。通过检测微滴内部的差异FRET发射并与已知结构进行比较，可以识别病毒RNA在各种条件下的结构变化。此外，高通量筛选可以鉴定特异性聚集并破坏病毒浓缩物的潜在抗病毒药物。更好地理解病毒相分离，沿着我将从这个项目中获得的技能，可以应用于其他感染系统。当面临抗病毒和抗微生物药物耐药性引起的全球危机时，这一点尤为重要。